Reengineering mRNA primary structure for enhanced protein production

a technology of enhanced protein and primary structure, which is applied in the direction of activity regulation, biochemistry apparatus and processes, and initiates by scanning mechanisms that do not consider the effects of potential initiation codons, and codons in mrna coding sequences and mirna-binding sites in mrnas that pose challenges to the pharmaceutical industry, etc., to achieve the effect of improving protein yield and concentration, improving efficiency and stability of protein translation

Active Publication Date: 2014-10-07
THE SCRIPPS RES INST
View PDF3 Cites 41 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0007]There is a need in the art for improving the efficiency and stability of protein translation and improving protein yield and concentration, for example, in the industrial production of protein drugs.

Problems solved by technology

However, the notion that translation initiates by a scanning mechanism does not consider the effects of potential initiation codons in coding sequences on protein synthesis.
The negative impact of putative initiation codons in mRNA coding sequences and miRNA-binding sites in mRNAs pose challenges to the pharmaceutical industry.
Poor protein expression limits the large-scale use of certain technologies, for example, problems in expressing enough antigen from a DNA vaccine to generate an immune response to conduct a phase 3 clinical trial.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reengineering mRNA primary structure for enhanced protein production
  • Reengineering mRNA primary structure for enhanced protein production
  • Reengineering mRNA primary structure for enhanced protein production

Examples

Experimental program
Comparison scheme
Effect test

example 1

Modification of Multiple Translation Initiation Sites within mRNA Transcripts

[0074]The presence of multiple translation initiation sites within the 5′-UTR and coding regions of mRNA transcripts decreases translation efficiency by, for example, diverting ribosomes from the authentic or demonstrated translation initiator codon. Alternatively, or in addition, the presence of multiple translation initiation sites downstream of the authentic or demonstrated translation initiator codon induces initiation of translation of one or more protein isoforms that reduce the translation efficiency of the full length protein. To improve translation efficiency of mRNA transcripts encoding commercially-valuable human proteins, potential translation initiation sites within all reading frames upstream and downstream of the authentic or demonstrated translation initiator codon are mutated to eliminate these sites. In preferred aspects of this method, the mRNA sequence is altered but the resultant amino ...

example 2

Modification of miRNA Binding Sites within mRNA Transcripts

[0099]MicroRNA (miRNA) binding to target mRNA transcripts decreases translation efficiency by either inducing degradation of the target mRNA transcript, or by preventing translation of the target mRNA transcript. To improve translation efficiency of mRNA transcripts encoding commercially-valuable human proteins, all known or predicted miRNA binding sites within a target mRNA's 5′ leader sequence, 5′ untranslated region (UTR) sequence, coding sequence, and 3′ untranslated region (UTR) sequence are first identified, and secondly mutated or altered in order to inhibit miRNA binding.

[0100]In a preferred aspect of this method, the seed sequence, comprising the first eight 5′- nucleotides of the mature miRNA sequence is specifically targeted. Seed sequences either include 5′ nucleotides 1-7 or 2-8 of the mature miRNA sequence. Thus, a seed sequence, for the purposes of this method, encompasses both alternatives. The miRNA seed seq...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
Login to view more

Abstract

Described herein are rules to modify natural mRNAs or to engineer synthetic mRNAs to increase their translation efficiencies. These rules describe modifications to mRNA coding and 3′ UTR sequences intended to enhance protein synthesis by: 1) decreasing ribosomal diversion via AUG or non-canonical initiation codons in coding sequences, and/or 2) by evading miRNA-mediated down-regulation by eliminating one or more miRNA binding sites in coding sequences.

Description

REFERENCE TO PRIORITY DOCUMENT[0001]This application claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Provisional Application Ser. No. 61 / 155,049, filed Feb. 24, 2009, entitled “Reengineering mRNA Primary Structure for Enhanced Protein Production.” The subject matter of the above-noted application is incorporated by reference in its entirety by reference thereto.INCORPORATION BY REFERENCE[0002]The contents of the text file named “37651-503001WO Sequence listing.txt” which was created on Oct. 26, 2011 and is 53 KB in size, are hereby incorporated by reference in their entirety.BACKGROUND[0003]Translation initiation in eukaryotes involves recruitment by mRNAs of the 40S ribosomal subunit and other components of the translation machinery at either the 5′ cap-structure or an internal ribosome entry site (IRES). Following its recruitment, the 40S subunit moves to an initiation codon. One widely held notion of translation initiation postulates that the 40S subunit moves from...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(United States)
IPC IPC(8): C12N15/11C12P21/02
CPCC12P21/02C12N15/111C12N2320/53C12N2320/50C12N2310/141C12P21/00C12N15/67C12N15/63
Inventor MAURO, VINCENT P.CHAPPELL, STEPHEN A.ZHOU, WEIEDELMAN, GERALD M.
Owner THE SCRIPPS RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap